Laboratory detection methods for the human coronaviruses

Shabani, Ehsan; Dowlatshahi, Sayeh; Abdekhodaie, Mohammad J.

doi:10.1007/s10096-020-04001-8

Cited by 24 publications

(26 citation statements)

References 131 publications

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“…PCR as a molecular technique is more suitable for accurate diagnoses because of its specificity in targeting and identifying specific pathogens [10]. Testing nucleotic acids amplification (NAAT) is the primary method of COVID-19 diagnoses and it has been recommended by the WHO for the better patients tests [11]. Samples are not only whole blood as described, but also respiratory samples (nasal and throat swabs), reliable 8 days after symptom onset.…”

Section: Clinical Application Of Pcr For Diagnosing Covid-19mentioning

confidence: 99%

“…SARS-CoV-2 virus has four structural proteins which can be identify with optimized PCR and RT-PCR protocols. The coronaviruses have a genome size of about 30 kilobases (kb) [11]. Several kits have been approved for detecting SARS-CoV-2 however, the reverse transcriptase PCR (RT-PCT) and the Real-Time Reverse transcriptase PCR (rRT-PCR) with respiratory samples are the most commonly method used for diagnoses to detect the virus genetically (Figure 2).…”

Section: Clinical Application Of Pcr For Diagnosing Covid-19mentioning

confidence: 99%

“…Several kits have been approved for detecting SARS-CoV-2 however, the reverse transcriptase PCR (RT-PCT) and the Real-Time Reverse transcriptase PCR (rRT-PCR) with respiratory samples are the most commonly method used for diagnoses to detect the virus genetically (Figure 2). Due to the rapid spread of coronavirus diseases a reliable detectable method is needed for an accurate diagnose [11]. The sequenced RNA of the virus which belongs to specific synthesis of proteins, is transcript into complementary DNA (cDNA) strands and subsequently the selected fragments of cDNA is amplified [9].…”

Section: Clinical Application Of Pcr For Diagnosing Covid-19mentioning

confidence: 99%

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Polymerase Chain Reaction (PCR) and How Temperature Modifies Nucleotide Pairings

Falgueras

2021

CTBEB

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Polymerase Chain Reaction (PCR) is a molecular technique more suitable for accurate diagnoses because of its specificity in targeting and identifying specific pathogens. PCR technique is reliable to detect the genetic presence of the virus in the host since the very beginning of the infection, no matter how the immune system of the person might react. Moreover, the samples for the test contains the DNA information during quite a long time since the extraction. Programmable thermal operation cycler for controlling heating and cooling is the procedure to denaturation into two single strands, annealing and extension reactions of the DNA. The PCR is certainly having a clinical approach, because do not emphasized on how the immune system of the patient might reacts to the infection, only on whether the specific virus is present or not for the accurate diagnoses and subsequently proceed with an appropriated medical treatment to restore health.

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Section: Clinical Application Of Pcr For Diagnosing Covid-19mentioning

confidence: 99%

Section: Clinical Application Of Pcr For Diagnosing Covid-19mentioning

confidence: 99%

Section: Clinical Application Of Pcr For Diagnosing Covid-19mentioning

confidence: 99%

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Polymerase Chain Reaction (PCR) and How Temperature Modifies Nucleotide Pairings

Falgueras

2021

CTBEB

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“…The early detection of SARS-COV-2 is necessary to control the widespread infection [ 139 – 160 ]. Several methods can be used for the diagnosis of COVID-19 [ 161 – 183 ]. The diagnosis of COVID-19 depends on the analysis of the patient's response due to the infection or the analysis of virus contents, e.g., RNA or their protein (Fig.…”

Section: Introductionmentioning

confidence: 99%

Nanobiotechnology as a platform for the diagnosis of COVID-19: a review

Abdelhamid

Badr

2021

Nanotechnol. Environ. Eng.

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A sensitive method for diagnosing coronavirus disease 2019 (COVID-19) is highly required to fight the current and future global health threats due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV 2). However, most of the current methods exhibited high false-negative rates, resulting in patient misdiagnosis and impeding early treatment. Nanoparticles show promising performance and great potential to serve as a platform for diagnosing viral infection in a short time and with high sensitivity. This review highlighted the potential of nanoparticles as platforms for the diagnosis of COVID-19. Nanoparticles such as gold nanoparticles, magnetic nanoparticles, and graphene (G) were applied to detect SARS-CoV 2. They have been used for molecular-based diagnosis methods and serological methods. Nanoparticles improved specificity and shorten the time required for the diagnosis. They may be implemented into small devices that facilitate the self-diagnosis at home or in places such as airports and shops. Nanoparticles-based methods can be used for the analysis of virus-contaminated samples from a patient, surface, and air. The advantages and challenges were discussed to introduce useful information for designing a sensitive, fast, and low-cost diagnostic method. This review aims to present a helpful survey for the lesson learned from handling this outbreak to prepare ourself for future pandemic.

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“…The SARS-CoV-2 genome consists of a single-stranded RNA molecule of approximately 30 kb in length [14]. Therefore, assays for detection of SARS-CoV-2 are often based on real-time reverse transcriptase polymerase chain reaction (RT-PCR) [15,16] and many RT-PCR assays have been given Emergency Use Authorization by the U.S. Food and Drug Administration [17]. The high analytical sensitivity of RT-PCR allows its use for pooled testing, in which multiple samples are mixed and tested in a single reaction, thus increasing testing e ciency and signi cantly lowering the costs [18].…”

Section: Introductionmentioning

confidence: 99%

Clinical evaluation of a multiplex RT-PCR assay for detection of SARS-CoV-2 in individual and pooled upper respiratory tract samples

Laverack

Tallmadge

Venugopalan

et al. 2021

Preprint

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The aim of this study was to identify and validate a sensitive, high-throughput and cost-effective SARS-CoV-2 RT-PCR assay to be used as a surveillance and diagnostic tool for SARS-CoV-2 in a University surveillance program. We conducted a side-by-side clinical evaluation of a newly developed SARS-CoV-2 multiplex assay (EZ-SARS-CoV-2 Real-Time RT-PCR) with the commercial TaqPath COVID-19 Combo kit, which has an Emergency Use Authorization from the FDA. The EZ-SARS-CoV-2 RT-PCR incorporates two assays targeting the SARS-CoV-2 N gene, an internal control targeting the human RNase P gene, and a PCR inhibition control in a single reaction. Nasopharyngeal (NP) and anterior nares (AN) swabs were tested as individuals and pools with both assays and in the ABI 7500 Fast and the QuantStudio 5 detection platforms. The EZ-SARS-CoV-2 RT-PCR assay analytical sensitivity was 250 copies/ml or approximately 1.75 genome copy equivalents per reaction. Clinical performance of the EZ-SARS-CoV-2 assay was determined using NP and AN samples tested in other laboratories. The diagnostic sensitivity of the assay ranged between 94 and 96% across the detection platforms, and the diagnostic specificity was 94.06%. The positive predictive value was 94% and the negative predictive value ranged from 94 to 96%. Pooling five NP or AN specimens yielded 93% diagnostic sensitivity. The overall agreement between these SARS-CoV-2 RT-PCR assays was high, supported by Cohen’s kappa value of 0.93. The EZ-SARS-CoV-2 RT-PCR assay performance attributes of high sensitivity, excellent performance in AN sample matrix and in pooled upper respiratory samples support its use in a high-throughput surveillance testing program.

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Laboratory detection methods for the human coronaviruses

Cited by 24 publications

References 131 publications

Polymerase Chain Reaction (PCR) and How Temperature Modifies Nucleotide Pairings

Polymerase Chain Reaction (PCR) and How Temperature Modifies Nucleotide Pairings

Nanobiotechnology as a platform for the diagnosis of COVID-19: a review

Clinical evaluation of a multiplex RT-PCR assay for detection of SARS-CoV-2 in individual and pooled upper respiratory tract samples

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